• Fisheries and Aquatic Sciences
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• v.26 no.2
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• pp.133-144
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• 2023

Auranofin is a US Food and Drug Administration (FDA)-approved anti-arthritis medication that functions as a thioredoxin reductase inhibitor. Spermidine, a polyamine present in marine algae, can exert various physiological functions. Herein, we examined the synergistic anticancer activity of auranofin and spermidine in hepatocellular carcinoma (HCC). Combined treatment with auranofin and spermidine suppressed cell viability more efficiently than either treatment alone in HCC Hep3B cells. The isobologram plotted by calculating the half maximal inhibitory concentration (IC₅₀) values of each drug indicated that the two drugs exhibited a synergistic effect. Based on the analysis of annexin V and cell cycle distribution, auranofin and spermidine markedly induced apoptosis in Hep3B cells. Moreover, auranofin and spermidine increased mitochondria-mediated apoptosis by promoting mitochondrial membrane potential (Δψm) loss. Auranofin and spermidine significantly increased reactive oxygen species (ROS) production in Hep3B cells, and the blocking ROS suppressed apoptosis induced by spermidine and auranofin. In addition, auranofin and spermidine reduced the expression of phosphorylated phosphatidylinositol-3 kinase (PI3K) and protein kinase B (Akt), and PI3K inhibitor accelerated auranofin- and spermidine-induced apoptosis. Using ROS scavenger and PI3K inhibitor, we revealed that ROS acts upstream of auranofin- and spermidine-induced apoptosis. Collectively, our study suggests that combination treatment with auranofin and spermidine could afford synergistic anticancer activity via ROS overproduction and reduced PI3K/Akt signaling pathway.

• Korean Journal of Animal Reproduction
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• v.17 no.2
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• pp.165-171
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• 1993

While ornithine decarboxylase (ODC) is considered a key enzyme in the biosynthesis of polyamines, difluoromethylornithine(DFMO) acts as an inhibitor of polyamine synthesis. Cycling crossbred gilts were randomly assigned to one of two (treatment and control) groups (6/group). An indwelling silicone catheter was surgically implanted in the jugular vein of each animal. DFMO was dissolved in saline(200 mg/ml) and adminstered by i. m. injection at a dose of 80 mg/kg/day. The control group received an equivalent volume saline injection. DFMO was injected 3 times daily(08:00. 16:00. 24:00h) from day 16 of estrous cycle to 21 or until estrus. Once daily blood samples (10ml) were taken from day 14 until two days after the last DFMO treatment. Window blood samples were collected every 15 min for 8 h (from 08:00 to 16:00h) starting on day 16 and continuing until day 21 from one gilt per day. Serum progesterone (P$_4$), estradiol (E$_2$), LH and FSH were measured. Typical concentration profiles for P$_4$ and E$_2$ were seen during the follicular phase regardless of DFMO treatment. Injection of DFMO suppressed the preovulatory LH concentration in the serum(p<0.01) while having no effect on FSH profile. The present results indicate that DFMO had an inhibitory effect on LH secretion in the pig, but did not affect PI, E2 or FSH release.

• Journal of Plant Biotechnology
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• v.38 no.1
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• pp.87-93
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• 2011

S-Adenosylmethionine decarboxylase (SAMDC; EC 4.1.4.50), a key enzyme for polyamines biosynthesis, was tightly regulated for homeostatic levels. Carnation SAMDC gene (CSDC9) has an small upstream open reading frame (uORF) of 54 amino acids in 5'-leader sequence. To explore the functional mechanism of uORFs in controlling translation, we used a GUS reporter gene driven with the 35S promoter and uORF region of SAMDC gene for making transgenic tobacco plants. In our experiment, there were a translational inhibition of its downstream GUS ORF by SAMDC uORF sequence or SAMDC uORF protein. Expecially, translational inhibition was most effective in point-mutated construct, in which the start codon was changed. Therefore, this results suggested the ribosomal stalling might be involved in this translational inhibitory process. The frame shift in amino acid sequence of SAMDC uORF with start codon and stop codon resulted in a moderate increasing in GUS activity, suggesting the native amino acid sequence was important for a function as a translational inhibitor. Also, we showed that the production of GUS protein was significantly inhibited in the presence of the small uORF using histochemical analysis of GUS expression in seedlings and tobacco flowers. Importantly, the small uORF sequence induced a real peptide of 5.7 kDa, which was provided the presence of SAMDC uORF peptide band using an in vitro transcription/translation system. The peptide product of uORF might interact with other components of translational machinery as well as polyamines, which was resulted from that polyamine treatment was inhibited GUS protein band in SDS-PAGE experiment.

• Journal of Plant Biotechnology
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• v.38 no.1
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• pp.54-61
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• 2011

Antisense construct of cDNA for senescencerelated ACC oxidase (CAO) cDNA isolated from carnation flowers were introduced into tobacco by Agrobacteriummediated transformation. The decreasing expression of NtACO and the reduction of ethylene production were observed in these transgenic lines. In contrast, the SAMDC transcripts and spermidine content were increased. The findings that higher content of spermidine in the ethylene suppressed transgenic plants compared with wild-type should be directly resulted in the enhancement of SAMDC activity followed by the increased accumulation of SAMDC transcript. To investigate the pathogenic response in these transgenic plants, wild-type and transgenic plants were inoculated with Phytophthora parasitica pv. nicotianae. Transgenic plants suppressing ethylene production showed the increased resistance against fungal pathogen, comparing with wild-type plant. PR-protein genes expression in CAO-AS-2 and CAOAS-4 were also higher at the normal growth condition and pathogenic response than in wild-type plants. The results of higher spermidine content and SAMDC activity in transgenic plants, CAO-AS-2 and CAO-AS-4, support the possibility that an increase in spermidine content might induce the higher transcripts of PR-protein genes. This results agreed with the phenomena that spermidine promoted the expression of PR1a and a SAMDC inhibitor, MGBG, decreased the expression of PR1a in leaf discs. These results suggest that the resistance against fungal pathogen in transgenic tobacco impaired in ethylene production might be caused by increasing in polyamine, especially spermidine, biosynthesis.

• Journal of Ginseng Research
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• v.44 no.3
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• pp.490-495
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• 2020

Background: Ginsenoside Rk1, a saponin component isolated from heat-processed Panax ginseng Meyer, has been implicated in the regulation of antitumor and anti-inflammatory activities. Although our previous studies have demonstrated that ginsenoside Rg3 significantly attenuated the activation of NMDA receptors (NMDARs) in hippocampal neurons, the effects of ginsenosides Rg5 and Rk1, which are derived from heat-mediated dehydration of ginsenoside Rg3, on neuronal NMDARs have not yet been elucidated. Methods: We examined the regulation of NMDARs by ginsenosides Rg5 and Rk1 in cultured rat hippocampal neurons using fura-2-based calcium imaging and whole-cell patch-clamp recordings. Results: The results from our investigation showed that ginsenosides Rg3 and Rg5 inhibited NMDARs with similar potencies. However, ginsenoside Rk1 inhibited NMDARs most effectively among the five compounds (Rg3, Rg5, Rk1, Rg5/Rk1 mixture, and protopanaxadiol) tested in cultured hippocampal neurons. Its inhibition is independent of the NMDA- and glycine-binding sites, and its action seems to involve in an interaction with the polyamine-binding site of the NMDAR channel complex. Conclusion: Taken together, our results suggest that ginsenoside Rk1 might be a novel component contributable to the development of ginseng-based therapeutic treatments for neurodegenerative diseases.

• The Korean Journal of Pharmacology
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• v.28 no.2
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• pp.115-128
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• 1992

The training of male wistar rats for active conditioned response (ACR) was performed by one daily training session of 30 consecutive trials for 10 successive days using a two-way shuttle box, and the rats that showed 10 or more ACRs on the last day were treated for further 10 days with electroconvulsive shock (ECS : 50 mA, 0.5 msec; 100 Hz; 1.5 sec) and the following compounds. On the 20th day, all the rats were tested for the ACR rention. The ECS regimens were one ECS per day for 10 days with one day interval $(5{\times}ECS)$, one ECS at 3 hrs (ECS-3h), and one ECS at 24 hrs (ECS-24h), respectively, before the ACR retention test. And CDP-choline (cc: 250 mg/kg), spermine (SM: 10 mg/kg), ${\alpha}-difluoromethylornithine$ (DO: 250 mg/kg), or aminoguanidine (AG: 100 mg/kg) was administered by one daily i.p. injection for 10 days. The ACR number $(13.7{\pm}1.0)$ obtained on the last training day was increased by 37.23% on the 20th day in the control rats. And the ACR increase was significantly suppressed by 5-ECS, ECS-3h, CC, or SM but was little affected by ECS-24h, DO, or AG. However, the 5-ECS induced impairment of ACR retention was significantly suppressed by AG, SM, and CC in the order of potency but was little affected by DFMO. And the ECS-3h induced impairment was moderately worsened by SM or AG. The acetylcholine (ACh) of the rat hypothalamus (HT), hippocampus (HC), and entorhinal cortex (EC) was markedly increased by CC and moderately increased by SM, but little affected by ECS-3h, ECS-24h, DO, or AG. But $5{\times}ECS$ slightly increased the ACh content. The brain putrescine (Pt) content was significantly increased by AG and little affected by CC, SM, or DO. But the $5{\times}ECS$ markedy decreased the brain Pt content, and the decrease was significantly suppressed by CC, SM, or AG. CC induced the marked increases of the spermidine (Sd) and spermine (Sm) contents of all the areas. SM increased the Sd contents of all the areas and the EC-Sm content. DO decreased the brain Sd and Sm contents. And AG increased the HT-Sd content and the Sm contents of all the brain areas. The $5{\times}ECS$ induced decrease of the HC-Sm content was suppressed by CC, SM and AG. These results suggest that the improving effect of aminoguanidine on the $5{\times}ECS$ induced impairment of ACR retention may be ascribed in part to its activity as a diamine oxidase inhibitor.